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Biomedical subjects

T Romano

Publications and source records attributed to T Romano.

At least 19 recordsLinked to original sources

Benzo[a]pyrene cytotoxicity in right whale (Eubalaena glacialis) skin, testis and lung cell lines.

The North Atlantic right whale (NARW) is one of the most endangered great whales. The NARW population consists of only about 300 individuals and is reproducing at an insufficient rate. There is growing concern about the potential effects of environmental contaminants on the reproductive and overall health of NARW. High contaminant burdens can accumulate in tissues of great whales but toxicological studies of their effects are limited due to legal, logistical and ethical restrictions and specific in vitro models are critically needed. Cell lines from NARW skin and internal organs were previously created in our laboratory. In this study, skin, testis and lung primary fibroblast cell lines were exposed to benzo[a]pyrene (BP) as part of a multi-chemical toxicity testing project in NARW. Cells were exposed for 24-72 h to 10 nM-10 microM BP dissolved in dimethylsulfoxide. Cytotoxicity was measured with a clonogenic assay using standard methods. Some cytotoxicity was observed after 24 h, the highest concentration (10 microM BP) resulting in 77, 74 and 51 percent relative survival in testis, skin and lung cells, respectively, and indicating a higher cytotoxicity in the lung (p < 0.05). After 48 and 72-h exposure, 10 microM BP resulted in 24 and 3, 74 and 27, and 42 and 23 percent relative survival in testis, skin and lung cells, respectively. Cytotoxicity significantly increased with exposure time in all three tissues (p < 0.05 for skin and p < 0.01 for lung and testis), suggesting metabolic activation of BP in the three organs. Fibroblast cytotoxicity observed in the testis was higher than that observed either in the skin or lung after 48 h (p < 0.01) and was close to 100% after 72 h, warranting further investigation of the potential effects of PAHs on reproductive health.

Animals↗

Changes in expression of monoclonal antibody epitopes on laminin-5r induced by cell contact.

Laminin-5r is a basement membrane component that promotes rapid adhesion and hemidesmosome formation in epithelial cells. We raised monoclonal antibodies and identified their corresponding epitopes on the constituent chains of laminin-5r by western blotting. Using a combination of immunoprecipitation and ELISA assays, we determined that these epitopes are differentially exposed on two forms of the laminin-5r heterotrimer: soluble (passively adsorbed onto plastic) and cell-associated. Antibody 5C5 epitope is exposed on the cell-associated form, but not the soluble/passively adsorbed form of laminin-5r. Epitopes reactive with antibodies CM6, FM3, and TR1 are also preferentially exposed on cell-associated laminin-5r, such that reactivity of these antibodies with the cell-associated form is fourfold higher than with the soluble/passively adsorbed form in ELISA assays. Incubation of passively adsorbed laminin-5r with the human epithelial cell line SCC12 induced exposure of 5C5 and CM6, FM3, or TR1 epitopes. These data suggest that cells actively modify laminin-5r, perhaps during matrix assembly, and that the 5C5 epitope may serve as a marker for assembled laminin-5r matrix.

Animals↗

Capture assay for specific IgE. An improved quantitative method.

A capture assay for the measurement of specific IgE in the serum of allergic patients is described, using monoclonal anti-human IgE (coated to the wells of a microtiter plate) and biotinylated allergens in solution. In a single incubation, IgE is bound to the solid phase through the Fc fragment and biotinylated allergens react with their specific IgE Fab regions, if present. In a second step, streptavidin-HRP conjugate is added to reveal the amount of biotin fixed on the solid phase. Quantitative determinations are obtained by comparison with a standard curve of total IgE incubated with a biotinylated monoclonal anti-IgE, complementary to the one employed as capture antibody. The assay is unaffected by allergen-specific IgG, shows good intra- and interassay reproducibility and is linear over a wide range of specific IgE concentrations. The method could be used with a wide range of different allergens.

Animals↗

Comparison of morphine tolerance in body temperature and luteinizing hormone secretion.

Small doses of morphine induce a hyperthermic response that does not change with repeated injections, whereas higher doses induce a hypothermia that changes to hyperthermia with repeated treatment. Experiment 1 confirmed these results in ovariectomized rats, using 6 repeated injections of morphine at 5 or 30 mg/kg. Using the same treatment regimen. Experiment 2 showed that the low dose of morphine induced a transient suppression of LH levels followed by a hypersecretion, and that repeated injections did not affect this response. The high dose (30 mg/kg) of morphine initially induced a sustained suppression of LH. Following the sixth injection of the high dose, subjects showed an intact or exaggerated suppression of LH, but an accelerated recovery to control levels. Experiment 3 extended the phase of repeated treatment of the high dose to 12 days and replicated the results of Experiment 2.

Animals↗

Immunohistochemical phenotyping of human solid tumors with monoclonal antibodies in devising biotherapeutic strategies.

A panel of 14 monoclonal antibodies (MoAbs) (4 raised against breast cancer, 6 against colon cancer and 4 against melanoma) were used to phenotype frozen sections of tumor biopsies obtained from 110 patients, by avidin-biotin-peroxidase complex techniques. We observed heterogeneity of antigen expression among the multiple metastatic lesions of single patients, as well as among tumor lesions from different patients with similar tumor histotypes. A wide range of cross-reactivity of anti-(breast-carcinoma) and anti-(colon-carcinoma) MoAbs with other carcinoma histotypes and limited reactivity with melanoma and sarcoma was detected. Some of our anti-melanoma MoAbs were also found to cross-react with selected carcinomas. Nine of the 14 MoAbs most reactive with carcinomas of diverse histotypes have been identified. A mixture or 'cocktail' of different MoAbs could be selected for each individual patient in order to achieve binding of MoAbs with most, if not 100% of tumor cells. This study illustrates the approach that we have taken to individualize the cocktail of MoAbs for the development of patient-specific therapeutic immunoconjugates.

Adult↗

An improved method for detecting and quantifying cardiac muscle disarray in hypertrophic cardiomyopathy.

The presence of muscle disarray in hypertrophic cardiomyopathy (HC), although well established, has only been semiquantitatively assessed. A quantitative method that uses polarized light microscopy is described. Hematoxylin and eosin-stained sections of ventricular septa from six HC patients and six normal hearts were examined. Cell orientations were measured in five regions from each section using a polarizing microscope with a rotating stage. Histograms of cell orientation angles were plotted and the mean and angular deviation of each sample were calculated. In normal hearts, cells were predominantly aligned parallel to each other. Orientation distributions were sharply peaked, with angular deviations ranging from 4 to 13 degrees. For HC, the sharp peak was lacking and angular deviations varied from 7 to 37 degrees; some distributions were bimodal. Areas in HC septa that appeared normal by gross inspection had abnormal orientation distributions. Polarized light microscopy provides an improved method of detecting and quantifying cellular disorganization in HC.

Cardiomyopathy, Hypertrophic↗

Transfer of delayed hypersensitivity in the chicken.

Delayed hypersensitivity (DH) can be transferred in FP and SC chickens with 1 x 10(8) spleen or peripheral blood cells from donors exhibiting moderate to strong DH reactions. Increasing the spleen cells dose from 1 to 4 x 10(8) per recipient had no marked enhancing effect on the results. Spleen cells from agammaglobulinemic donors were also shown to transfer DH reactions.

Animals↗

Gallium-67-citrate scanning in tuberculous peritonitis.

A 32-year old black, alcoholic male was noted to have an abnormal Gallium-67-citrate scan during a work-up for persistent fever. Surgical biopsies revealed findings of the dry form of tuberculosis peritonitis. The literature concerning Gallium-67-citrate scanning was reviewed and its usefulness in inflammatory disease states (including two reports of positive scans in tuberculous peritonitis) was demonstrated. In addition, we have considered the possible mechanisms of Gallium-67-citrate uptake by inflammatory cells and its probable relationship to the granulomas associated with tuberculous peritonitis. We conclude that Gallium-67-citrate scanning is an important diagnostic procedure in the evaluation of a fever of unknown origin. Moreover, it may significantly shorten the diagnostic work-up of those cases of tuberculous peritonitis.

Adult↗